US 7,407,805 B2 · Skin substitutes with improved barrier function

Abstract

The present invention relates to in vitro cultured skin substitutes, and in particular to in vitro cultured skin substitutes that have improved barrier function. In some embodiments, improved barrier function is a result of improved culture conditions, while in other embodiments, improved barrier function results from genetic modification of keratinocytes. Improved culture conditions to improve barrier function include organotypic culture in the presence of linoleic acid and/or linoleic acid at about 75% humidity. Suitable genetic modifications for improving barrier function includes transfection with a DNA construct capable of expressing GKLF.

Claims7

Dependent claims

A method of making a skin equivalent having improved barrier function, the method comprising:

providing (a) a composition comprising near-diploid immortalized keratinocyte (NIKS) cells and (b) a culture medium comprising ascorbic acid, linoleic acid, isoproterenol, carnitine, serine, oleic acid, arachidonic acid, α-tocopherol, and epidermal growth factor in amounts effective to improve barrier function;
and
culturing said composition (a) in said culture medium (b) under conditions wherein said culturing provides a skin equivalent having a surface electrical capacitance of 40 to 240 pF measured over a 10 second interval in vitro.

Dependencies are inferred from the claim text.

Facts

Application

11/235,814Filed 27 Sep 2005 · division

Priority

2 Mar 2001

Published

US 2006/0121608 A18 Jun 2006

Granted

5 Aug 2008

Term ends

Not estimatedThe source records don’t support an estimate.

Classification15 codes

C12N 5/0698
Artificial constructs associating cells of different lineages, e.g. tissue equivalents: skin equivalents
C12N 2500/36
Organic components: lipids
C12N 2500/38
Organic components: vitamins
C12N 2500/90
Specific components of cell culture medium: serum-free medium, which may still contain naturally-sourced components

Family

Application60/273,034Us provisional application · filed 2 Mar 2001

Application10/087,346Parent of this division · filed 1 Mar 2002

US 2006/0121608 A1Pre-grant publication · published 8 Jun 2006

US 6,974,697 B2Parent of this division

US 2002/0168768 A1Published application

US 7,407,805 B2This patent

From the US filing history. Foreign family members aren’t listed.

Skin substitutes with improved barrier function

Life of this patent

Skin substitutes with improved barrier function

Life of this patent

Abstract

Claims7